title: Development of enzyme-linked immunosorbent assay (ELISA) for glutathione S-transferase (GST-S) protein in the intertidal copepod Tigriopus japonicus and its application for environmental monitoring.
abstract: To utilize the GST-S protein as a useful biomarker for environmental contamination, we developed a polyclonal antibody-based enzyme-linked immunosorbent assay (ELISA) in the intertidal copepod Tigriopus japonicus. Two polyclonal antibodies, TJ-GST-S1 and TJ-GST-S2, were raised against two TJ-GST-S synthetic peptides. Also a recombinant TJ-GST-S protein was purified as a standard for ELISA development. Each polyclonal antibody was tested by Western blot analysis and indirect ELISA. Of two polyclonal antibodies, TJ-GST-S2 ELISA was further employed due to its wide range of detection and the limit of specificity compared to those of TJ-GST-S1 ELISA system. After exposure to 4 metals (Ag, As, Cd, and Cu) to T. japonicus, the amount of TJ-GST-S protein was significantly elevated in a concentration-dependent manner. Also, TJ-GST-S protein was upregulated at relative high concentrations of B[α]P, PCB, and TBT. In this paper, we suggest that T. japonicas ELISA for TJ-GST-S2 is useful as a potential indicator system for marine contaminants.
authors.affiliation: Research Institute for Natural Sciences, Hanyang University, Seoul 133-791, South Korea.
authors.forename: Jae-Sung
authors.initials: JS
authors.lastname: Rhee
authors.forename: Bo-Mi
authors.initials: BM
authors.lastname: Kim
authors.forename: Chang-Bum
authors.initials: CB
authors.lastname: Jeong
authors.forename: Kenneth Mei Yee
authors.initials: KM
authors.lastname: Leung
authors.forename: Gyung Soo
authors.initials: GS
authors.lastname: Park
authors.forename: Jae-Seong
authors.initials: JS
authors.lastname: Lee
mesh_terms: D000818:Animals; D033342:Copepoda / Q000378:metabolism*; D004784:Environmental Monitoring / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D015870:Gene Expression; D005982:Glutathione Transferase / Q000235:genetics / Q000378:metabolism*; D014874:Water Pollutants, Chemical / Q000633:toxicity*
chemical_list: D014874:Water Pollutants, Chemical; D005982:Glutathione Transferase
keywords: Contaminant; Copepod; Enzyme-linked immunosorbent assay; Glutathione S-transferase; Tigriopus japonicus